CV17-Zero Background pTOPO-Blunt Simple Cloning Kit（零背景平末端克隆，不含多克隆MCS）

產(chǎn)品介紹：

       本制品和傳統(tǒng)的T4連接酶原理不同，它利用了Topoisomerase可以在瞬間（幾秒鐘-幾分鐘）、高效（接近100%）連接DNA片段的原理采用本公司獨(dú)創(chuàng)的工藝制成。 

1. 平末端PCR產(chǎn)物不需要先加A，在瞬間（幾秒鐘-幾分鐘）完成平末端PCR產(chǎn)物連接。 

2. 特制的新型載體質(zhì)粒大小僅僅不到2kb，充分發(fā)揮了TOPO載體越小，可容納片段越大的優(yōu)勢，最大限度提高了大片段連接效率；連接后質(zhì)粒大小比傳統(tǒng)載體小2kb以上，質(zhì)粒越小，轉(zhuǎn)化效率越高，極大的提高了各種片段連接后的轉(zhuǎn)化子數(shù)量。 

3. 采用氨芐抗性載體只需10分鐘復(fù)蘇時間，比卡那抗性載體1小時復(fù)蘇時間縮短6倍。 

4. 最快可以不用冰浴和熱休克，室溫5分鐘內(nèi)完成轉(zhuǎn)化；無需1小時復(fù)蘇，只需37℃10分鐘復(fù)蘇便可以涂板。從連接到涂板最快只需15-20分鐘。

5. 自殺基因零背景原理，無自連假陽性，無需繁瑣藍(lán)白斑篩選和菌落PCR篩選。大部分情況下隨機(jī)挑一個克隆便是有插入的（接近100%）。

6. 連接長片段能力遠(yuǎn)超傳統(tǒng)Blunt克隆載體，可連接長達(dá)10kb片段（例如連接5kb片段，也可能達(dá)到挑10個菌落，至少8個是有插入的效果），是新一代世界領(lǐng)先的簡單、快速、零背景免篩選的TOPO Blunt平末端克隆載體。

本制品在克隆插入位點(diǎn)兩側(cè)不含多克隆酶切位點(diǎn)（Simple），需要時可在PCR擴(kuò)增引物上導(dǎo)入合適的酶切位點(diǎn)。此時如果使用PCR擴(kuò)增引物導(dǎo)入的酶切位點(diǎn)進(jìn)行酶切時，酶切反應(yīng)將不會受到載體上其它多克隆酶切位點(diǎn)影響，可大大提高酶切效率，增加亞克隆成功率。

注：本TOPO載體連接體系是按照10ml計(jì)算可以用20次，如果按照其它公司產(chǎn)品5ml計(jì)算，實(shí)際使用次數(shù)可以增加一倍，20次實(shí)際可用40次。 

使用該產(chǎn)品發(fā)表的部分文章：

CV17 Zero Background pTOPO-Blunt Simple Cloning Kit::

1. Cloning and Expression Analysis of Citrate Synthase(CS)Gene in Camellia oleifera. Bulletin of Botanical Research 2016, 36(4): 556-564

2. Cloning and expression analysis of a autophagy-related protein 8 encoding gene in Polyporus umbellatus. Chin Med Biotechnol 2016, 11(5): 415-419

3. Cloning and expression of three thaumatin-like protein genes from Polyporus umbellatus. Acta Pharmaceutica Sinica B 2017, 7(3): 373–380

4. Development and application of an allele-specific PCR assay for detecting T409C mutation of cyp51 gene linked with tebuconazole resistance in Villosiclava virens (rice false smut)  Canadian Journal of Plant Pathology 2017, 39(3)

5. LEA proteins from Gastrodia elata enhance tolerance to low temperature stress in Escherichia coli. Gene 2018, 646: 136-142

6. Complete genomic sequence of a novel macluravirus, alpinia oxyphylla mosaic virus (AloMV), identified in Alpinia oxyphylla. Archives of Virology 2018,  https://doi.org/10.1007/s00705-018-3879-62

7. Complete nucleotide sequence of jasmine virus H, a new member of the family Tombusviridae. Archives of Virology 2018, 163(3): 731-735

8. Analysis of the complete genome sequence of a potyvirus from passion fruit suggests its taxonomic classification as a member of a new species. Archives of Virology 2018, https://doi.org/10.1007/s00705-018-3885-8

9. Endogenous pararetrovirus sequences are widely present in Citrinae genomes. Virus Research 2018, https://doi.org/10.1016/j.virusres.2018.05.018

10. Molecular cloning, polymorphism, and expression analysis of the LKB1/STK11 gene and its association with non-specific digestive disorder in rabbits. Molecular and Cellular 2018,  https://doi.org/10.1007/s11010-018-3349-1